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Thromb Haemost 1992; 68(05): 600-605
DOI: 10.1055/s-0038-1646324
Original Article
Schattauer GmbH Stuttgart

Quantitation of Platelet Glycoprotein IV (CD36) in Healthy Subjects and in Patients with Essential Thrombocythemia Using an Immunocapture Assay

Valérie Thibert
1   The INSERM U 353, Hôpital St. Louis, Paris, France
,
Sylvia Bellucci
2   The Département d'Angio-Hématologie, Hôpital Lariboisière, Paris, France
,
Lena Edelman
3   The Institut Pasteur, Paris, France
,
Narendra N Tandon
4   The American Red Cross, Cell Biology Laboratory, Rockville, MD, USA
,
Chantal Legrand
1   The INSERM U 353, Hôpital St. Louis, Paris, France
› Author Affiliations
Further Information

Publication History

Received 05 March 1992

Accepted after revision 22 June 1992

Publication Date:
04 July 2018 (online)

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Summary

Glycoprotein IV (GPIIIb, CD36) is a major platelet membrane glycoprotein which is thought to participate in a number of adhesive reactions and to mediate signal transduction. In order to measure the total content of GPIV in human platelets, we have developed a simple and sensitive solid-phase radioimmunoassay based on the immunocapture of GPIV from Triton X-100-solubilized platelets. FA6-152, a monoclonal antibody to GPIV was coated on microtiter plates and bound antigen was quantified with a radiolabeled polyclonal antibody to GPIV Using purified GPIV as a standard, the coefficients of variation of the assay were found to be less than 10% at concentrations of GPIV ranging from 0.15 to 0.75 µg/ml. The assay was validated by the parallelism obtained between purified GPIV dose-response curves and those obtained with platelet lysates, indicating a similar antigenic activity for GPIV in both samples. The level of GPIV in platelets from healthy donors was 0.23 ± 0.05 (mean ± SD, n = 15) µg per 100 µg of platelet proteins and a mean value of 27,440 ± 6,200 (SD) molecules per platelet was calculated. The radioimmunoassay could be used to discriminate between the high level of platelet GPIV in patients with essential thrombocythemia (mean ± SD = 81,850 ± 27,780 molecules/platelet; n = 8) and the normal GPIV level in patients with secondary thrombocytosis (mean ± SD = 26,810 ± 4,030 molecules/platelet; n = 5), thereby demonstrating the clinical usefulness of the assay. The specific increase in platelet GPIV in patients with essential thrombocythemia was confirmed by immunoblot analysis whereas no increase in platelet GPIb or GPIIb-IIIa was observed by this technique.

 

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